Like others [4, 15] we also detected a strong up-regulation of SM

Like others [4, 15] we also BAY 11-7082 clinical trial detected a strong up-regulation of SMA positive cells in CDE livers. GW3965 datasheet Interestingly, periportal SMA positive cells co-expressed vimentin, a protein actually synthesized in fibroblasts [34], suggesting their origin

from periportal (myo-)fibroblasts rather than from HSCs, since co-expression of GFAP, a characteristic for the transdifferentiation into myofibroblasts demonstrated in vitro [35, 36] but not in vivo, was rarely detectable. Even though we might have missed such an event in an early phase after exposure to CDE, it is remarkably that the above mentioned activation of HSC persists even after two weeks. Thus, HSCs seem to have other functions than transdifferentiation to myofibroblasts as it was discussed in a recent study using a rat oval cell

model selleck compound [37]. Up-regulation of CD31 (PECAM) in livers of CDE treated mice is another new finding of this study. The lack of any BrdU/CD31 co-expression points to an increase of CD31 in SECs. In untreated livers CD31 positive cells were hardly detected, whereas up-regulation seems to be associated with dedifferentiation of SECs into a defenestrated endothel during pseudocapillarization due to fibrotic processes [38] which also occur under CDE conditions [4]. The impact of re-expression of LI-cadherin in adult mouse liver during CDE diet is still unclear and currently under investigation in double knock-out mice for LI and E-cadherin in our group. Possibly, re-expression of LI-cadherin, an embryonal marker of mouse liver [39], prevents the dissociation of cellular

connections on sites of insufficient expression of E-cadherin. Conclusions The present study clearly shows that in mouse liver M2-Pk is expressed in 2-hydroxyphytanoyl-CoA lyase nearly all cells of hepatic sinusoid. Undisputable CDE diet leads to an up-regulation of M-Pk, but this rise is the summation of M1- and M2-Pk. The elevation should no longer be misinterpreted as a specific oval cell response. Under CDE conditions GFAP expressing cells expand in a zonal specific pattern. Pericentral GFAP positive cells seem to present an activated cell type. Periportal oval cells express GFAP, a common HSC marker. Therefore, this marker does not seem suitable for tracing progenitors of hepatocytes under CDE conditions. Methods Animals GFAP-tTA mice (B6.Cg.Tg(GFAP-tTA)110Pop/J, Jacksons Laboratory, Bar Harbor, USA) were intercrossed with ptetCre mice (LC1, [40]) resulting in double transgenic mice expressing Cre-recombinase by GFAP promoter driven tTA expression (GFAP-Cre-mice). Mice of mixed genetic backround (DAB/C57Bl/6) and GFAP-Cre mice were given a CDE diet over 14 days. Cholin deficient animal chow without addition of methionine (Altromin, Lage, Germany) was provided ad libitum and drinking water was replaced by 0.165% ethionine solution (TCI, Europe, Zwijndrecht, Belgium) and was also given ad libitum.

elgii B69, in which at least 5 NRPS-related

biosynthetic

elgii B69, in which at least 5 NRPS-related

biosynthetic gene clusters were found within its 7,981,270 bp long scaffold [11]. Further inspection revealed that several NRPS genes located in scaffolds 3 and 43 were probably related with pelgipeptin biosynthesis. The gaps between and within these two scaffolds were filled by sequencing PCR products. These efforts resulted in a complete NRPS gene cluster (plp), harbouring eight open reading frames (ORFs), which could be assigned to pelgipeptin biosynthesis. These ORFs (designated plpA-plpH) were transcribed in the same direction (Figure1B). Upstream of the plp locus, two genes (ORF2 and ORF3) encoding proteins with similarities to heparinase II/III family proteins

learn more (YP_003243728 and YP_003243727, respectively) were transcribed in the same direction and were considered not to be involved in pelgipeptin production. Further upstream, a third ORF (ORF1), with TGA stop codon within ORF2, was found to encode a protein with high similarity to short-chain dehydrogenases/reductases (ZP_08509633) and was also considered not involved in the pelgipeptin biosynthesis. Downstream of the plpF gene, four genes encoding putative ABC transporter proteins were found. PlpG and PlpH, shared 72% and 69% identities with PmxC and PmxD, respectively, which were considered MM-102 responsible for the secretion of polymyxin produced by P. polymyxa[12]. This transport activity may be needed for the transport of pelgipeptin out of the cell, Thiamet G and therefore, the gene products were attributed to pelgipeptin biosynthesis. The other two genes (ORF4 and ORF5) encoding putative nitrate/sulphonate/bicarbonate ABC transporter proteins were transcribed in

the opposite direction and were considered less likely to be involved in pelgipeptin production, although further evidence will be required before this can be decided unequivocally. The putative ORFs and the genetic organisation of the chromosomal region containing these sequences are depicted in Figure1B. Genes encoding NRPS As shown in Figure1B, three NRPS genes, plpD plpE, and plpF, are present in the plp cluster, and these genes encode proteins with estimated molecular masses of 171.8, 951.3, and 122.9 kDa, respectively. The modules and selleck compound domains of pelgipeptin synthetase were analysed as described in the “Materials and methods” section above. PlpD, containing four domains (C-A-T-C) (Figure1B), had an N-terminal C domain, which shared 43% identity with the starter C domain of PmxE [12]. The amino acid predicted specific for the A domain of PlpD was 2,4-diaminobutyric acid (Dab) (Table1). The presence of a starter C domain in PlpD, and the specificity of the module for Dab are both consistent with this module providing the first amino acid of the pelgipeptin peptide, and therefore the fatty acid side chain should be connected to the peptide at this residue [13].

A limitation of this study was that it did not investigate the po

A limitation of this study was that it did not investigate the potential effects of omeprazole on IPE through CYP2C19 inhibition or change in gastric pH, although this is not expected. Another limitation was the relatively short study duration, given the potentially long duration of use of either one of the study drugs alone or when used concomitantly, although typically, drug–drug interaction studies are relatively short in duration. AZD6738 purchase 5 Conclusions At steady state, IPE

4 g/day did not inhibit the AUC0–24 and C max of the CYP2C19 substrate omeprazole at 40 mg/day. Coadministration of these two drugs was safe and well tolerated in this PK study of healthy adult subjects. Acknowledgments This study was designed and sponsored by Amarin Pharma Inc., Bedminster,

NJ, USA. Medical writing assistance was provided by Beth Daro-Kaftan, PhD, of Peloton Advantage, LLC, Parsippany, NJ, USA, and funded by Amarin Pharma Inc. Declaration of interest Dr. Stirtan is an employee and stock shareholder of Amarin Pharma Inc. Drs. Braeckman and Soni are former employees and current stock shareholders of Amarin Pharma Inc. Open AccessThis article is distributed under the terms of the AZD4547 Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original 4SC-202 cell line author(s) and the source are credited. References 1. Ford ES, Li C, Zhao G, Pearson WS, Mokdad AH. Hypertriglyceridemia and its pharmacologic treatment among US adults. Arch Intern Med. 2009;169:572–8.PubMedCrossRef 2. Third Report of the National Cholesterol Education Baf-A1 molecular weight Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) final report. Circulation. 2002;106:3143–421. 3. Berglund L, Brunzell JD, Goldberg AC, Goldberg IJ, Sacks F, Murad MH, Stalenhoef AF. Evaluation and treatment of hypertriglyceridemia: an endocrine society clinical practice guideline. J Clin Endocrinol Metab. 2012;97:2969–89.PubMedCentralPubMedCrossRef 4. Vascepa [package insert]. Bedminster: Amarin Pharma Inc.; 2013.

5. Bays HE, Ballantyne CM, Kastelein JJ, Isaacsohn JL, Braeckman RA, Soni PN. Eicosapentaenoic acid ethyl ester (AMR101) therapy in patients with very high triglyceride levels (from the Multi-center, plAcebo-controlled, Randomized, double-blINd, 12-week study with an open-label Extension [MARINE] trial). Am J Cardiol. 2011;108:682–90.PubMedCrossRef 6. Ballantyne CM, Bays HE, Kastelein JJ, Isaacsohn JL, Braeckman RA, Soni PN. Efficacy and safety of eicosapentaenoic acid ethyl ester (AMR101) therapy in statin-treated patients with persistent high triglycerides (from the ANCHOR study). Am J Cardiol. 2012;110:984–92.PubMedCrossRef 7. Yacyshyn BR, Thomson AB. The clinical importance of proton pump inhibitor pharmacokinetics. Digestion. 2002;66:67–78.PubMedCrossRef 8.

Hygrophoroideae tribe Chrysomphalineae Our 4-gene backbone analy

Hygrophoroideae tribe Chrysomphalineae. Our 4-gene backbone analyses, however, show strong support for placing Cantharellula in subf. Lichenomphalioideae rather than Hygrophoroideae. Nevertheless, subfamilies Lichenomphalioideae and Hygrophoroideae, are in adjacent clades, so the appearance of similar hymenial architecture in both clades suggests a possible homologous origin. Fig. 20 Subf. Lichenomphalioideae, tribe Cantharelluleae, Pseudoarmillariella ectypoides lamellar cross section (DJL05, North

Carolina, Kinase Inhibitor Library order Great Smoky Mt. National Park, USA). Scale bar = 20 μm Fig. 21 Subf. Lichenomphalioideae, tribe Cantharelluleae, Cantharellula umbonata lamellar cross section (RDY-1366, R. Youst, California, USA). Scale bar = 20 μm Tribe Cantharelluleae is the only group retained in the Hygrophoraceae with amyloid spores. Neohygrophorus angelesianus check details (A.H. Sm. & Hesler) Singer (= Hygrophorus subg. Pseudohygrophorus A.H. Sm. & Hesler) is shown as sister to Tribe Clitocybeae (Tricholomataceae) in a multigene Supermatrix analysis by Matheny

et al. (2006), sister to the type of Pseudoomphalina, P. kalchbrenneri, (in the Tricholomataceae), in our 4-gene backbone analyses (100 % MLBS; 1.0 BPP), and sister to Pseudoomphalina felloides in previous Supermatrix (Lodge et al. 2006) and LSU analyses (Moncalvo et al. 2002; 70 % MPBS). Another species with amyloid spores, Hygrophorus metapodius old (Fr.) Fr. [≡Camarophyllus metapodius (Fr.) Wünsche, ≡Hygrocybe metapodia (Fr.) M.M. Moser, ≡Neohygrocybe metapodia (Fr.) Herink], was also transferred to the Tricholomataceae and recombined in gen. Porpoloma by Singer (1973). Pseudoarmillariella ectypoides has been

variously placed in Clitocybe (Saccardo 1887), Clitocybula (Raithelhuber 1980) and Omphalina (Bigelow 1982), while Cantharellula has been placed in Cantharellus (Persoon 1794), and Hygrophoropsis (Kühner and Romagnesi 1953). Singer (1942; 1948; 1986) recognized the close Selleck AZD0156 relationship between Cantharellula umbonata and Pseudoarmillariella ectypoides, but placed them together with other amyloid spored genera in the Tricholomataceae, tribe Leucopaxilleae. Singer transferred Peck’s Agaricus ectypoides to Cantharellula in 1942, erected subg. Pseudoarmillariella Sing. in 1948 for C. umbonata and C. ectypoides (Peck) Singer, then raised subg. Pseudoarmillariella to genus rank for P. ectypoides in 1965. Moncalvo et al. (2002) were the first to show inclusion of tribe Cantharelluleae in the Arrhenia–Lichenomphalia clade (as cantharelloid clade 62) using an LSU analysis, but without significant branch support. Using a four-gene Supermatrix analysis, Lodge et al. (2006) were the first to show significant support for the Cantharelluleae clade, while Matheny et al. (2006) were the first to show significant Bayesian support (1.0 PP) for including Pseudoarmillariella in the Hygrophoraceae and subf. Lichenomphalioideae.

Astrophys J 249:481–503CrossRef Córdova A, Engqvist M, Ibrahem I,

Astrophys J 249:481–503CrossRef Córdova A, Engqvist M, Ibrahem I, Casas J, Sunde´n H (2005) Plausible origins of homochirality in the amino acid see more catalyzed neogenesis of carbohydrates. Chem Commun 2047–2049 Córdova A, Zou W, Dziedzic P, Ibrahem I, Reyes

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Thus, discrimination between C1 and C2 statements was based on ex

Thus, discrimination between C1 and C2 statements was based on expert consensus. 5. GSK872 Publication and future revisions The Guidelines were published in the Japanese-language journal of the JSN and concurrently released as a Japanese-language book (by Tokyo Igakusha, Tokyo). The Guidelines were also uploaded to the homepage of the JSN. At

present, CKD-related evidence is being rapidly accumulated, and this new evidence will necessitate the preparation of an updated version of the Guidelines in 3–5 years. A certain degree of turnover in the membership of the revision committee will be required in order to ensure the impartiality of the Guidelines.”
“Introduction LY2874455 clinical trial Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used, with acknowledged efficacy and safety over a wide range of clinical conditions. Despite their many useful therapeutic applications, substantial evidence has shown that NSAIDs can have deleterious effects

on kidney function. For example, a nested case-controlled study using the General Practice Research Database from the United Kingdom showed that NSAID users in the general population were at threefold greater risk for a first-ever diagnosis of clinical acute kidney injury (AKI) than non-NSAID users. In addition, history of heart failure, hypertension, and diabetes were associated with a greater risk of AKI in this population [1]. Combination therapy with NSAIDs and renin–angiotensin system (RAS) inhibitors increases the risk of kidney damage [2–4]. Since RAS inhibitors are recommended as first-line antihypertensive agents in patients with diabetes, patients this website with diabetic nephropathy who take NSAIDs tend to be at greater risk for NSAID-induced kidney damage. NSAIDs can affect renal function

by, for example, inhibiting the synthesis of important renal prostaglandins, especially those involved in solute homeostasis and maintenance of renal Inositol oxygenase blood flow [5–8]. Prostaglandin E2 (PGE2) is the most abundant vasodilatory prostaglandin in the human renal vascular bed. NSAIDs decrease PGE2 concentration by inhibiting cyclooxygenase-2 (COX-2). Adverse effects of NSAIDs may be avoided by administering these drugs as transdermal patches. These adhesive patches, which are applied to the skin at the site of pain, slowly release medication through the skin. Although NSAID patches are regarded as safe and are frequently used in patients with chronic kidney disease (CKD), the effects of NSAID patches on renal circulation in these patients have not been investigated. Loxoprofen-containing patches are one of the most widely used adhesive patches in Japan. We therefore analyzed the effects of topically applied loxoprofen sodium on kidney function in patients with diabetic nephropathy. Methods Study design This open-label, single-arm, single-dose study was performed at the Shiga University of Medical Science Hospital.

Then, the mixture was shifted into a dialysis membrane (MWCO of 3

Then, the mixture was shifted into a dialysis membrane (MWCO of 3,000) learn more against pure water to remove surplus PEG2000N. Characterization To determine the size and morphology, RNase A@C-dots were characterized by high-resolution transmission electron microscopy (HR-TEM, JEM-2100 F, 200 kV, JEOL Ltd., Tokyo, Japan). The samples for TEM/HR-TEM were made by simply dropping

aqueous solution of the C-dots onto a 300-mesh copper grid casted with a carbon film. UV–Vis absorption spectra of the C-dots were measured with a Varian Cary 50 LY3039478 spectrophotometer (Varian Inc., Palo Alto, CA, USA). Fluorescence excitation and emission spectra of RNase A@C-dots were recorded on a Hitachi FL-4600 spectrofluorimeter (Hitachi Ltd., Tokyo, Japan). Zeta potential of RNase A@C-dots was measured on a Nicomp 380 ZLS zeta potential/particle sizer (PSS. Nicomp, Santa Barbara, CA, USA). X-ray photoelectron

spectroscopy (XPS) was obtained at room temperature by a Kratos Axis Ultra spectrometer selleck chemical (AXIS-Ultra DLD, Kratos Analytical Ltd., Tokyo, Japan) using a monochromated Al Kα (1486.6 eV) source at 15 kV. Fourier transform infrared (FTIR) spectra were obtained on a Nicolet 6700 spectrometer (Thermo Electron Corporation, Madison, WI, USA). The samples for FTIR measurement were prepared by grinding the dried C-dots with KBr together and then compressed into thin pellets. X-ray diffraction (XRD) profiles of the C-dot powders were recorded on a D/MAX 2600 PC (Rigaku, Tokyo, Japan) equipped with graphite monochromatized Cu Kα (λ = 0.15405 nm) radiation at a scanning speed of 4°/min in the range from 10° to 60°. Time-resolved fluorescence intensity decay of RNase A@C-dots was performed on a LifeSpec II (Lifetime only, Edinburgh Instruments, Livingston, UK). The sample was excited

by 380-nm laser, and the decay was measured in a time scale of 0.024410 ns/channel. Quantum yield measurement To assess the quantum yield of RNase A@C-dots, quinine sulfate in 0.1 M H2SO4 (quantum yield, 54%) was used as a reference fluorescence reagent. The final results were calculated according to Equation 1 below: (1) where Φstd is the known quantum yield of the standard compound, F sample and F std stand Tideglusib for the integrated fluorescence intensity of the sample and the standard compound in the emission region from 380 to 700 nm, A std and A sample are the absorbance of the standard compound and the sample at the excitation wavelength (360 nm), and n is the refractive index of solvent (for water, the refractive index is 1.33). To minimize the reabsorption effects, UV absorbance intensities of the samples and standard compound should never exceed 0.1 at the excitation wavelength. Photoluminescence (PL) emission spectra of all the sample solutions were measured at the excitation wavelength of 360 nm. The integrated fluorescence intensity is the area under the PL curve in the wavelength from 380 to 700 nm.

frequency is extracted

and shown in the inset of Figure 6

frequency is extracted

and shown in the inset of Figure 6. Strong frequency dispersion is observed for all of the samples. It is clear that the deteriorative degree of dielectric relaxation increases from 12.1 nm, reaches the peak at 22.5 nm, and then declines. A comparison between the samples of 12.1 and 25 nm is made. Uniformly, the sample with the grain size of 25 nm is shown to perform superior on dielectric relaxation. The dielectric constant frequency response of the PNZT samples shares exactly the same response for the CeO2 samples (one dielectric relaxation peak within the frequency range). A possible reason [19] to the cited observation could be the broadened dielectric peak and the transition temperature shift. The dielectric constant shows phase transition as expected for normal ferroelectrics. The region around the dielectric peak is broadened, which is one of the most important characteristics of disordered perovskite structure with the diffuse phase ABT-737 mouse transition. The transition temperature is found to shift forward to lower temperature with the grain size from 12.1 to 22.5 nm, while the transition 4EGI-1 cost temperature remains at the same position with further increasing grain size. Concerning the strong frequency dispersion, it is mainly

attributed to the low-frequency space charge accumulation effect. Such strong frequency dispersion in dielectric constant appears to be a common feature in ferroelectrics associated with non-negligible ionic conductivity. Therefore, the reason for the

dielectric relaxation of the PNZT samples could be the possible mechanism behind the frequency dependence of the k value of the CeO2 samples. Many dielectric relaxation models (Cole-Davidson, Havriliak-Negami, and Kohlrausch-Williams-Watts) were proposed to interpret the dielectric relaxation, which is also termed as the frequency dependence of the k value. The Havriliak-Negami (HN) model is suitable Glycogen branching enzyme for almost all of the high-k materials as it has three parameters for fitting (α, β, and τ). In contrast, the Cole-Davidson (CD) model only has two parameters for fitting (β and τ). Thus, if the CD model is able to fit the cerium oxides, it will be more significant for the specified physical mechanism compared to the HN model. Concerning the Kohlrausch-Williams-Watts (KWW) model, it has also two adjusting parameters for fitting (β and τ). The CD and KWW models have certain links in both high frequency and low frequency approximations. Selleck Daporinad Besides, the CD model is widely used in glass-forming materials to explain the frequency dependence of the dielectric constants [20]. Here, dielectric relaxation can be described by the CD law for all of the CeO2 samples. CD fittings are denoted by solid lines in Figure 6. In 1951, D. W. Davidson and R. H. Cole [21] proposed the CD equation to interpret data observed on propylene glycol and glycerol based on the Debye expression. The CD equation can be represented by ϵ*(ω).

[5, 32] (Figure 6a) At the same time, the PL component peaked at

[5, 32] (Figure 6a). At the same time, the PL component peaked at 700 to 750 nm can be attributed to the defects located at Si-nc/MK5108 order matrix interface because slight increase of its maximum magnitude is apparently due to overlapping with

near-infrared component which intensity increases with cooling (Figure 6a, curve 3). Based on the PL results, one can conclude that the main contribution to the PL spectra in our samples is given by the carrier recombination through different defects. The high concentration of interface and matrix defect (in particular, the high intensity of PL band at 700 to 750 nm) obviously hinders the observation of exciton recombination. Conclusions Selleckchem OSI-027 The effect of annealing treatment on structural and light emission properties of Si phase-rich Al2O3 films with different Si contents was investigated. The formation of amorphous Si clusters upon deposition process was observed for the films with x ≥ 0.38. The annealing results in the formation check details of Si crystallites whose mean size depends on the type of post-deposition treatment. The conventional annealing of the samples with

x = 0.5 to 0.68 causes the formation of Si-ncs with the mean size of about 14 nm, whereas similar samples submitted to rapid thermal annealing show the presence of Si-ncs with sizes of about 5 nm. Two main broad PL bands were observed in the 500- to 900-nm spectral range with peak positions at 575 to 600 nm and 700 to 750 nm as well as near-infrared tail. The low-temperature measurements revealed that the first PL band was unchanged with cooling, while the slight increase of maximum intensity of the second one was obviously due to overlapping with near-infrared band. Such behavior of visible PL bands differs from that expected for quantum

confined Si-ncs that allowed ascribing them to interface and/or matrix defects. At the same time, the analysis of PL spectrum shape allows ascribing the near-infrared PL component (780 to 900 nm) to the exciton recombination inside Si-ncs. Acknowledgments This work was supported by the National Academy of Sciences of Ukraine, Ministry of Art and Science of Israel. One of the authors (LK) would like to acknowledge also the French National Research Agency for partial financial support. References 1. Canham LT: Silicon quantum wire array fabrication Protein kinase N1 by electrochemical and chemical dissolution of wafers. Appl Phys Lett 1990, 57:1046–1048.CrossRef 2. Lehman V, Gosele U: Porous silicon formation: a quantum wire effect. Appl Phys Lett 1991, 58:856–858.CrossRef 3. Shimizu-Iwayama T, Nakao S, Saitoh K: Visible photoluminescence in Si + -implanted thermal oxide films on crystalline Si. Appl Phys Lett 1994, 65:1814–1816.CrossRef 4. Chen XY, Lu YF, Tang LJ, Wu YH, Cho BJ, Xu XJ, Dong JR, Song WD: Annealing and oxidation of silicon oxide films prepared by plasma-enhanced chemical vapor deposition.

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Cond Res 2003,17(4):822–831.PubMed 28. Volek JS, Ratamess NA, Rubin MR, Gomez AL, French DN, McGuigan MM, Scheett TP, Sharman MJ, Hakkinen K, Kraemer WJ: The effects of creatine supplementation on muscular performance and body composition responses to short-term resistance training overreaching. Eur J Appl Physiol 2004,91(5–6):628–637.PubMedCrossRef 29. Kingsley M, Cunningham D, Mason L, Kilduff LP, McEneny J: Role of creatine supplementation on exercise-induced cardiovascular function and oxidative stress. Oxid Med Cell Longev 2009,2(4):247–254.PubMedCentralPubMedCrossRef 30. Deminice R, Rosa FT, Franco GS, Jordao AA, de Freitas EC: Effects of creatine supplementation on oxidative stress and inflammatory markers after repeated-sprint exercise in humans. Microtubule Associated inhibitor Nutrition 2013,29(9):1127–1132.PubMedCrossRef 31. Deminice R, Jordao AA: Creatine supplementation www.selleckchem.com/products/BKM-120.html reduces oxidative stress biomarkers after acute exercise in rats. Amino Acids 2012,43(2):709–715.PubMedCrossRef 32. Botezelli JD, Cambri LT, Ghezzi AC, Dalia RA, M Scariot PP, Ribeiro C, Voltarelli FA, Mello MA: Different exercise protocols improve metabolic syndrome markers, tissue triglycerides content and antioxidant status in rats. Diabetol Metabol Syndr 2011, 3:35.CrossRef 33. Lambertucci RH, Levada-Pires AC, Rossoni LV, Curi R, Pithon-Curi TC: Effects of aerobic exercise training on antioxidant enzyme activities and mRNA levels in soleus muscle from young and aged rats.